Ultra-performance Liquid Chromatography-electrospray Ionization Research Articles

Transcription and Metabolism Pathways of Anthocyanin in Purple Shamrock (Oxalis triangularis A.St.-Hil.).

Anthocyanins are water-soluble pigments that can impart various colors to plants. Purple shamrock (Oxalis triangularis) possesses unique ornamental value due to its purple leaves. In this study, three anthocyanins, including malvidin 3-O-(4-O-(6-O-malonyl-glucopyranoside)-rhamnopyranosyl)-5-O-(6-O-malonyl-glucopyranoside), delphinidin-3-O-rutinoside and malvidin-3,5-di-O-glucoside, were characterized with ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) in purple shamrock. To investigate the molecular mechanism of anthocyanin biosynthesis in green shamrock (Oxalis corymbosa) and purple shamrock, RNA-seq and qRT-PCR were performed, and the results showed that most of the anthocyanin biosynthetic and regulatory genes were up-regulated in purple shamrock. Then, dark treatment and low temperature treatment experiments in purple shamrock showed that both light and low temperature can induce the biosynthesis of anthocyanins.

Enhancement of Digestion Resistance and Glycemic Control of Corn Starch through Conjugation with Gallic Acid and Quercetin Using the Free Radical Grafting Method

The objective of this study was to synthesize different polyphenol–corn starch complexes including gallic acid–starch and quercetin–starch by conjugating corn starch with gallic acid and quercetin using the free radical grafting method. This process was effective in enhancing conjugations of starch molecules with gallic acid and quercetin (5.20 and 5.83 mg GAE/g, respectively) and imparted promising antioxidant capacity to the phenolic–starch complexes. Significant interactions between these phenolic compounds and corn starch molecules were revealed with an ultraperformance liquid chromatography electrospray ionization Q-time-of-flight mass spectrometry assay. It was revealed that significantly higher levels of resistant starch in the above gallic–starch and quercetin–starch complex samples (11.6 and 15.3 g/100 g, respectively) together with an obvious reduction in glycemic response (7.9% and 11.8%, respectively) observed over the control. Complex samples functionalized with gallic acid and quercetin have exerted modified physicochemical properties, particularly reduction in swelling ability (58.7–60.1%), breakdown viscosity (62.5–67.8%), and setback viscosity (37.7–44.5%). In sum, free radical grafting treatment could be a promising method for imparting corn starch with enhanced resistance to enzyme digestion along with changes in pasting properties for specific food applications.

Lipidomic analysis and triglyceride profiles of fish oil: Preparation through silica gel column and enzymatic treatment

A comprehensive study of lipidomic coupled with triglyceride profiles onto four fish oils was performed through ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). Overall, 1010 lipids belonging to 6 categories and 38 lipid classes were identified. Triglycerides (TGs) were the dominant component in four fish oils (40 %–99 % of total lipids), and glycerophospholipids (GPs) and sphingolipids (SLs) were another two major lipid categories in the fish oil (TG50) which prepared through silica gel column. These results revealed that enzymatic treatment has slight effect on lipid distribution but silica gel column could change the lipids composition. TGs composition of four fish oils were separated completely, and the most TG molecule in TG50 is TG(18:3_14:0_18:0), possessed 13.03 ± 5.07 % relative content, these results implied that silica gel column could protect the nature structure of TGs from destroying which may also limited to further improve eicosapentaenoic acid/docosahexaenoic acid (EPA/DHA) purity, but enzymic method was not restricted by this.

Nontarget Metabolites of Rhizomes of Edible Sacred Lotus Provide New Insights into Rhizome Browning

The “edible rhizome” variant of Nelumbo nucifera with various cultivars has a long history of use as a food in East Asia. In this study, 48 target metabolites were untargeted and identified in 212 rhizome cultivars (tropical and temperate types) using ultraperformance liquid chromatography-electrospray ionization quadrupole time-of-flight high-resolution mass spectrometry; among these, 32 compounds were newly reported in the rhizome. Combined with the browning phenotype of 212 lotus rhizomes, (epi) catechin, norarmepavine, and N-feruloyl-3-methoxytyramine were used as predominant chemical markers to separate different degrees of lotus rhizome browning. p-Coumaroyltyramine and N-trans-feruloyltyramine were selected as predominant chemical markers to investigate the differential expression between tropical and temperate lotus using principal component analysis and orthogonal partial least squares discriminant analysis. Shared and unique structure plots were used to compare the outcomes of the ecotype and browning OPLS model, showing that variation in tropical lotus rhizome browning is not obvious; this will be of great importance for genetic improvement by providing a hereditary basis.

Determination of the Oxidative Stress Biomarkers of 8-Hydroxydeoxyguanosine and Dityrosine in the Gills, Skin, Dorsal Fin, and Liver Tissue of Atlantic Salmon (Salmo salar) Parr.

Oxidative stress is a condition caused by an imbalance in the occurrence of reactive oxygen species in the cells and tissues of organisms. An ultra-performance liquid chromatography–electrospray ionization tandem mass spectrometry (UPLC–ESI–MS/MS) method was developed for the simultaneous determination of two oxidative stress biomarkers, 8-hydroxydeoxyguanosine (8OHDG) and dityrosine (DIY), in the gills, skin, dorsal fin, and liver tissue of Atlantic salmon (Salmo salar) parr. The use of target analyte-specific 13C and 15N internal standards allowed quantification of each target analyte to be performed through the standard solvent calibration curve. The relative recoveries [mean ± (relative standard deviation%)] of 8OHDG and DIY were 101 ± 11 and 104 ± 13% at a fortified concentration of 10 ng/mL (8OHDG) and 500 ng/mL (DIY), respectively, ensuring the accuracy of the extraction and quantification. The chromatographic separation was carried out using a gradient elution program with a total run time of 5 min. The limits of detection (LODs) were 0.11 and 1.37 ng/g wet weight (w.w.) for 8OHDG and DIY, respectively. To demonstrate the applicability of the developed method, it was applied in 907 tissue samples that were collected from Atlantic salmon parr individuals reared in an experimental land-based recirculating aquaculture system (RAS) treated with peracetic acid. Moreover, the possibility of using the dorsal fin as an alternative matrix for the minimally invasive assessment of oxidative stress in Atlantic salmon parr was introduced. To our knowledge, 8OHDG and DIY were used for the first time as biomarkers for biomonitoring the fish health (oxidative stress) of Atlantic salmon parr in RAS.

Lipidomic profile changes of yellow-feathered chicken meat during thermal processing based on UPLC-ESI-MS approach

In this work, ultra-performance liquid chromatography-electrospray ionization mass spectrometry (UPLC-ESI-MS) was applied to compare the lipidomics profile of chicken breast meat heated to various temperatures. A total of 445 lipids including phospholipids, diacylglycerols, triacylglycerols, glycoglycerolipids, and sphingolipids were detected in chicken muscle. Significant decreases in a list of characteristic unsaturated phospholipid species including phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylinositol were identified after heating, suggesting the occurrence of lipid oxidation, while significant increases in characteristic lysophospholipid species were found with increasing heating temperature, which were mainly derived from hydrolysis of phospholipids. Moreover, ether phospholipid and neutral lipid species exhibited remarkable changes during the heating process, and monoalk(en)yl diacylglycerol was first identified in chicken meat. The findings could contribute to the improved understanding of key lipids and biochemical reactions engaged in the heating of meat products.

Effect of fermentation stages on glucosinolate profiles in kimchi: Quantification of 14 intact glucosinolates using ultra-performance liquid chromatography-tandem mass spectrometry.

We developed and validated an ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry-based analytical method to determine intact glucosinolates in kimchi and evaluate the effects of fermentation stages on glucosinolate profiles. The developed method yielded reliable data in the kimchi matrix in terms of selectivity, matrix effect (88%-105%), linearity (coefficients of determination ≥0.9991), sensitivity (limits of quantification ≤35nmol/L), accuracy (82%-101%), and precision (≤8%). The kimchi samples contained progoitrin, sinigrin, glucoraphanin, glucoraphenin, glucoalyssin, gluconapin, glucobrassicanapin, glucobrassicin, glucoberteroin, gluconasturtiin, 4-methoxyglucobrassicin, and neoglucobrassicin, of which 4-methoxyglucobrassicin, glucobrassicanapin, and gluconapin were the major compounds. Total glucosinolate content was decreased by 31%-97% and 91-100% in the moderate-fermented and over-fermented samples, respectively, compared with that in the non-fermented samples, revealing sudden glucosinolate degradation between the moderate- and over-fermentation stages. In summary, we report an efficient analytical method to estimate kimchi glucosinolate profiles, which could be a foundation for future studies.

Mahuang Xixin Fuzi decoction ameliorates apoptosis via the mitochondrial-mediated signaling pathway in MCM cells

Ethnopharmacological relevanceMahuang Xixin Fuzi Decoction (MXF), as a classical prescription of traditional Chinese medicine (TCM), has been used to treat the symptoms of fever, nasal congestion and headache in elderly people for almost a thousand years. Aim of the studyThe purpose of this study was to evaluate the effects and possible mechanisms of MXF on thermal stimulation-induced mouse cardiac myocytes (MCM) cell apoptosis. Materials and methodsThe apoptosis of the MCM cell model was induced by a PCR-calculated temperature control system with a gradual heating pattern at 43 °C for 1 h. The cytotoxic effects were determined using real-time cell analyzer (RTCA) technology. Annexin V-FITC/7-AAD staining, and JC-1 fluorescence were used to assess apoptosis. Specific substrates, enzyme-linked immunosorbent assays (ELISAs), and Western blotting were used to identify proteins in the mitochondrial-mediated pathway. The identification of chemical components in the mouse heart was performed by ultra-performance liquid chromatography–electrospray ionization tandem mass spectrometry analysis. ResultsMXF inhibited apoptosis through the mitochondrial-mediated signaling pathway, including ameliorating ∆Ψm reduction, blocking mitochondrial Cyt C release, reducing Bax levels and increasing Bcl-2 levels, suppressing caspase-9 and caspase-3 activation in cytoplasmic fractions. Moreover, the components of MXF that act on the heart are mainly ephedra alkaloids and aconitine alkaloids. ConclusionsThe findings demonstrated that MXF treatment markedly reduced MCM cell apoptosis induced by thermal stimulation, which may be ascribed to the mitochondrial-mediated signaling pathway.

Enhanced Antioxidant, Antiviral, and Anticancer Activities of the Extract of Fermented Egyptian Rice Bran Complexed with Hydroxypropyl-β-cyclodextrin.

Egyptian rice bran was fermented with baker’s yeast, and released phenolics were extracted with aqueous methanol to give fermented rice bran extract (FRBE). The analysis of the FRBE with ultra-performance liquid chromatography–electrospray ionization–tandem mass spectrometry revealed 21 compounds, mainly phenolic acids and flavonoids. The FRBE was then complexed with (2-hydroxypropyl)-β-cyclodextrin (HPβCD) via noncovalent host–guest inclusion complexation using the thin-film hydration technique to improve the hydrophilicity and bioactivity of the FRBE. The formation of the inclusion complex was confirmed using HPLC, 1H NMR, FT-IR, and a phase solubility study. In addition, the biological activities of the complex were investigated. The FRBE/HPβCD inclusion complex had more pronounced antioxidant, antiviral, and anticancer activities compared to free FRBE. These findings warrant the future investigation of potential medical applications of FRBE.

Green fast and simple UPLC-ESI-MRM/MS method for determination of trace water-soluble vitamins in honey: Greenness assessment using GAPI and analytical eco-scale

Honey is one of the most commonly used natural products for nutritional and therapeutic purposes. Besides sugars, honey is characterized by complex chemical composition of minerals, polyphenols, water soluble vitamins, carotenoids and organic acids. In the present study, nine water soluble vitamins; C (L-ascorbic acid), B1 (thiamine), B2 (Riboflavin), B3H (nicotinic acid), B5 (D-pantothenic acid), B6 (pyridoxine), B7 (Biotin), B9 (Folic acid) and B12 (Cyanocobalamin) are determined simultaneously for the first time using an ultra-performance liquid chromatography-electrospray ionization multiple reaction monitoring/mass spectrometry (UPLC-ESI-MRM/MS) method coupled with a single simple sample preparation step. The simplicity and sensitivity of the proposed method permit the simultaneous determination of all nine water soluble vitamins including vitamins B7 and B12 that are present in trace amounts in various real honey samples. Complete chromatographic separation is achieved in 3 mins on a C18 column with a green mobile phase composed of a mixture of aqueous formic acid (0.1%, v/v) and methanol in gradient elution mode at a flow rate of 0.4 mLmin−1. The method is fully validated according to ICH guidelines. The greenness of our proposed method is assessed by comparing it to other proposed methods in terms of sample preparation, instrument energy consumption, consuming hazardous reagents and waste generation using Analytical Eco-scale and GAPI (Green Analytical Procedure Index). Results proved the superior greenness of our proposed method over the other reported ones. The proposed method is applied to twelve samples from six different monofloral honey types. Quantitation of the water-soluble vitamins content revealed concentrations ranging from 0.63 to 936.98 mgKg−1 in the analyzed honey samples with the successful determination of vitamins B7 and B12 in trace amounts. The highest vitamin content detected was for vitamin B3 in Jujube-tree honey while the lowest vitamin content was for vitamin B12 in Orange-tree honey. A marked relation was observed between the botanical origin and the vitamins’ concentrations of the studied honey samples. In addition, factors such as temperature, moisture, light and pH during storage greatly affect the vitamins content of honey. The developed method is intended to serve as a reliable, accurate, fast and green tool for the routine analysis of water-soluble vitamins in honey samples for quality control and research purposes.

Influence of ripening stage and meteorological parameters on the accumulation pattern of polyphenols in greengages (Prunus mume Sieb. Et Zucc) by widely targeted metabolomic

Greengage is a Chinese traditional medicine food homology plant that contains abundant polyphenols. Greengages with different levels of maturity have different medicinal functions and applications. Therefore, this study comprehensively analysed the phenolic compounds in greengage of whole mature stage. Notably, the influence of climate conditions on greengage polyphenol synthesis was deeply explored. The polyphenols of greengages were profiled based on the widely targeted metabolomic technology using ultra-performance liquid chromatography–electrospray ionisation triple quadrupole mass spectrometry (UPLC–QTRAP–MS/MS). A total of 214 polyphenols were detected, covering 11 subclasses. During the ripening of greengages, these polyphenols first increased and then decreased, peaking during the swell stage. Multivariate statistical methods, including redundancy analysis (RDA), random forest analysis, Mantel test, and Spearman's correlation indicated that temperature, sunshine hours, humidity, and radiation were important factors driving the formation and changes in the polyphenols of greengages. In particular, flavanones and flavonols, showed a structure-dependent response to temperature and radiation.

Effects of Fengycins and Iturins on Fusarium oxysporum f. sp. physali and Root Colonization by Bacillus velezensis Bs006 Protect Golden Berry Against Vascular Wilt.

Bacillus velezensis Bs006 has shown antagonistic activity on Fusarium oxysporum f. sp. physali and biocontrol activity against Fusarium wilt (FW) in golden berry (Physalis peruviana). We hypothesized that strain Bs006 has the ability to synthesize antimicrobial cyclic lipopeptides (CLPs) like other members of the same species. However, if so, the real effects of CLPs on F. oxysporum f. sp. physali and their potential as a biocontrol tool against Physalis-FW have not been elucidated. In this study the CLPs profile of Bs006 in liquid culture and antagonist-plant-pathogen interactions were characterized. Also, the potential effects of supernatant free of bacteria against F. oxysporum f. sp. physali and FW were explored and compared with the effects of pure CLPs. Ultraperformance liquid chromatography-electrospray ionization-mass spectrometry analysis revealed the capacity of Bs006 to synthesize homologous compounds of iturins, surfactins, and fengycins in liquid culture and on the inhibition zone against F. oxysporum f. sp. physali in dual confrontation tests. Bs006 supernatant reduced the germination and growth of F. oxysporum f. sp. physali and caused vacuolization, swelling, and lysis of F. oxysporum f. sp. physali cells in a concentration-dependent manner. Pure fengycins affected the development of F. oxysporum f. sp. physali from 11 mg/liter and iturins from 21 mg/liter. In a gnotobiotic system, Bs006 colonized the root surface of golden berry, inhibited the growth of F. oxysporum f. sp. physali, and produced CLPs. Individual application of Bs006 and supernatant protected the plants from F. oxysporum f. sp. physali infections by 37 to 53%, respectively. Meanwhile, fengycins reduced the disease progress by 39%. These results suggest further studies to select an optimum combination of Bs006 and supernatant or CLPs, which might be a good option as biofungicide against F. oxysporum f. sp. physali.

Effect of oridonin on oxylipins in the livers of mice with acute liver injury induced by D-galactosamine and lipopolysaccharide

Background and PurposeOridonin (Ori) has been shown to protect against acute liver injury (ALI) induced by D-galactosamine (D-GalN) and lipopolysaccharide (LPS). Oxylipins are oxidation products of polyunsaturated fatty acids (PUFAs) and are key proinflammatory mediators. This study aimed to investigate the changes in oxylipins in the livers of mice with D-GalN/LPS-induced ALI and the effects of Ori on these changes. Results54 oxylipins in liver tissues were identified and qualitatively and quantitatively analyzed by ultra-performance liquid chromatography-electrospray ionization triple quadrupole mass spectrometry (UPLC-QTRAP/MS/MS). The levels of 12-HETE, 12-HEPE, 14(S)-HDHA, PGE2, dihomo-γ-linolenic acid and 13-HOTrE in the liver were significantly increased in the D-GalN/LPS-induced ALI group compared with the control group, and the levels of EPA and 7-HDHA were significantly decreased. However, pretreatment with Ori dramatically decreased the levels of 12-HETE, 12-HEPE, 14(S)-HDHA, PGE2 and 13-HOTrE compared with those of the ALI group and induced 7-HDHA and 15-oxoETE. Moreover, Ori reduced the protein levels of COX-1, COX-2, ALOX5, ALOX12 and ALOX15 induced by D-GalN/LPS, indicating that Ori altered oxylipins through the COX and LOX pathways. ConclusionsThese results suggest that the protective effect of Ori on ALI is partly mediated by affecting the oxylipin pathway.

Enhancement on antioxidant and antibacterial activities of Brightwell blueberry by extraction and purification

A blueberry anthocyanin extract was obtained from Brightwell blueberry fruits cultivated in eastern China and the extraction and purification conditions were optimized. The components of the anthocyanin extract were identified using ultra-performance liquid chromatography-electrospray ionization interface-mass spectrometer. The antioxidant and antibacterial activities of the blueberry fruit supernatant (BFS), blueberry anthocyanin crude extract (BCE), and blueberry anthocyanin rich extract (BRE) were evaluated. The extraction yield was 1.79 ± 0.0014 mg/g under the following optimal conditions: 1:20 solid-to-liquid ratio (v/w), 24 h, 34 °C, and 90% ethanol containing 0.21% (v/v) hydrochloric acid. With regard to purification, anthocyanin purity increased 19.1-fold. Nine fractions were identified as the glycosides of delphinidin, cyanidin, petunidin, and malvidin. The biological activities of the blueberry anthocyanin extract were improved through extraction and purification. Compared with BFS and BCE, BRE had a higher DPPH radical scavenging activity (EC50 = 0.51 mg/mL), ABTS antioxidant capacity (EC50 = 0.32 mg/mL), and oxygen radical absorbance capacity (0.43 mmol Trolox/g). Furthermore, BRE (2 mg/mL) showed a maximum of 84.64 ± 0.35% reduction in the biofilm biomass of Listeria monocytogenes and the inhibition zone given by BRE against Escherichia coli was 16.04 ± 0.38 mm. BRE showed the highest antioxidant capacities and obvious antibacterial effects against food-related microorganisms than the other samples. Therefore, BRE can be used as a natural antioxidant and antibacterial agent and has potential health advantages and food industry applications.

Rapid Determination of Per- and Polyfluoroalkyl Substances (PFAS) in Harbour Porpoise Liver Tissue by HybridSPE®-UPLC®-MS/MS.

A rapid hybrid solid phase extraction (HybridSPE®) protocol tailored to ultra-performance liquid chromatography–electrospray ionization tandem mass spectrometry (UPLC®–ESI–MS/MS) analysis was developed for the determination of 15 per- and polyfluoroalkyl substances (PFAS) in liver tissue from harbour porpoises (Phocoena phocoena). The HybridSPE® technique has been applied in trace concentration bioanalysis, but it was mainly used for liquid biological media until now. In this study, the protocol was applied on tissue matrix, and it demonstrated acceptable absolute recoveries (%) ranging from 44.4 to 89.4%. The chromatographic separation was carried out using a gradient elution program with a total run time of 4 min. The inter-day method precision ranged from 2.15 to 15.4%, and the method limits of detection (LODs) ranged from 0.003 to 0.30 ng/g wet weight (w.w.). A total of 20 liver samples were analyzed to demonstrate the applicability of the developed method in liver tissue from a wildlife species.

Determination of SK7324, a new class of hepatitis C virus inhibitor, in rat plasma by UPLC-MS/MS and its application to the pharmacokinetic study of SK7324.

In this study, a sensitive method for quantitation of the unique small-molecule inhibitor of hepatitis C virus SK7324 in rat plasma has been established using ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI/MS/MS). SK7324 and internal standard (tramadol) in plasma sample was extracted using acetonitrile. A centrifuged upper layer was then evaporated and reconstituted with the mobile phase of 0.5% formic acid-acetonitrile (35:65, v/v). The reconstituted samples were injected into a C18 reversed-phase column. Using MS/MS in the multiple reaction monitoring (MRM) mode, SK7324 and tramadol were detected without severe interference from rat plasma matrix. Detection of SK7324 in rat plasma by the UPLC-ESI/MS/MS method was accurate and precise with a quantitation limit of 1.0 ng/mL. The validation, reproducibility, stability, and recovery of the method were evaluated. The method has been successfully applied to pharmacokinetic studies of SK7324 in rat plasma. Pharmacokinetic parameters of SK7324 were evaluated after intravenous (i.v.; at doses of 5 mg/kg) and oral (p.o.; at doses of 10 mg/kg) administration of SK7324 in rats. After p.o. administration (10 mg/kg) of SK7324, F (Fraction absorbed) value was approximately 87.1%.

Trihydroxycholanoyl-taurine in brains of rodents with hepatic encephalopathy.

Hepatic encephalopathy (HE), a neurological disease resulting from liver failure, is difficult to manage and its causes are unclear. Bile acids have been postulated to be involved in the provenance and progression of various diseases including HE. Hence, the characterization of bile acid profiles in the brains of subjects with and without liver failure can provide important clues for the potential treatment of HE. Nanoflow ultra-performance liquid chromatography electrospray ionization ion mobility mass spectrometry (UPLC-ESI-IM-MS) is a highly sensitive method for detection of specific molecules, such as bile acids in brain samples, at biologically relevant concentrations. We used UPLC-ESI-IM-MS to characterize bile acid profiles in brain samples from seven "healthy" control rodents and 22 "diseased" rodents with liver failure (i.e., induced HE). An isomer of trihydroxycholanoyl-taurine was detected in brain tissue samples from both rats and mice with induced HE; however, this isomer was not detected in the brains of healthy rats and mice. Our findings were confirmed by comparing IM arrival times (AT), exact mass measurements (m/z), and mass spectral fragmentation patterns of the experimentally observed suspected species to standards of trihydroxycholanoyl-taurine isomers. Moreover, In Silico Fractionation was employed to provide an additional analytical dimension to verify bile acid identifications.

Alteration in the Cerebrospinal Fluid Lipidome in Parkinson's Disease: A Post-Mortem Pilot Study.

Lipid metabolism is clearly associated to Parkinson’s disease (PD). Although lipid homeostasis has been widely studied in multiple animal and cellular models, as well as in blood derived from PD individuals, the cerebrospinal fluid (CSF) lipidomic profile in PD remains largely unexplored. In this study, we characterized the post-mortem CSF lipidomic imbalance between neurologically intact controls (n = 10) and PD subjects (n = 20). The combination of dual extraction with ultra-performance liquid chromatography-electrospray ionization quadrupole-time-of-flight mass spectrometry (UPLC-ESI-qToF-MS/MS) allowed for the monitoring of 257 lipid species across all samples. Complementary multivariate and univariate data analysis identified that glycerolipids (mono-, di-, and triacylglycerides), saturated and mono/polyunsaturated fatty acids, primary fatty amides, glycerophospholipids (phosphatidylcholines, phosphatidylethanolamines), sphingolipids (ceramides, sphingomyelins), N-acylethanolamines and sterol lipids (cholesteryl esters, steroids) were significantly increased in the CSF of PD compared to the control group. Interestingly, CSF lipid dyshomeostasis differed depending on neuropathological staging and disease duration. These results, despite the limitation of being obtained in a small population, suggest extensive CSF lipid remodeling in PD, shedding new light on the deployment of CSF lipidomics as a promising tool to identify potential lipid markers as well as discriminatory lipid species between PD and other atypical parkinsonisms.

CF–PK–PD analysis of natural hemostatic compounds from Toddalia asiatica (Linn) Lam root bark in rats

AbstractNatural hemostatic compounds from Toddalia asiatica (Linn) Lam (T. asiatica) root bark had been investigated by a novel strategy, chemical fingerprint–pharmacokinetic–pharmacodynamic (CF–PK–PD) for the first time in this study. The extract sample of T. asiatica root bark was subdivided into petroleum ether (PE), ethyl acetate (EA) and n-butanol (n-B) sample by reagent extraction, EA sample showed significant hemostatic activity using prothrombin time (PT), activated partial thromboplastin time (APTT) and fibrinogen (FIB) as evaluation indexes from rat plasma of PK experiment in hemorrhagic rat model. CF analysis was adopted to assist us to discover six natural compounds from T. asiatica root bark in actual rat plasma after sample treatment by Ultra Performance Liquid Chromatography-Electrospray Ionization (UPLC-ESI) MS, there were only lomatin and 5-methoxy-8-hydroxy psoralen showing significant hemostatic effect (P < 0.05) mainly through endogenous coagulation pathway and fibrinolytic system. In PK–PD study, six compounds in EA sample exhibited relatively rapid absorption and slow elimination characteristics. The mean Tmax and t1/2β of isopimpinellin and pimpinellin were 1.74 and 0.59 h, 5.31 and 6.89 h in rats. On the basis of Sigmoid–Emax model, PK–PD related curves of FIB in hemorrhagic rat model after treatment of T. asiatica root bark were obtained. Predicted Emax, EC50 and ke0 of FIB under isopimpinellin were 4.87 mg/mL, 1.39 μg/mL and 0.81 1/h; predicted Emax, EC50 and ke0 of FIB under pimpinellin were 4.29 mg/mL, 2.47 μg/mL and 0.77 1/h. In conclusion, hemostatic compounds from T. asiatica root bark had been materialized, there were lomatin, isopimpinellin, pimpinellin and 5-methoxy-8-hydroxy psoralen at least as its main active substances through coagulation pathways and fibrinolytic system. CF–PK–PD method as a promising method was worthy of follow-up opening, application in pharmaceutical research.

Peganum harmala Alkaloids Self-Assembled Supramolecular Nanocapsules with Enhanced Antioxidant and Cytotoxic Activities.

Amphiphilic macrocycles, such as p-sulfonatocalix[6]arenes (p-SC6), have demonstrated great potential in designing synthetic nanovesicles based on self-assembly approaches. These supramolecular nanovesicles are capable of improving the solubility, stability, and biological activity of various drugs. In the present study, the biologically active harmala alkaloid-rich fraction (HARF) was extracted from Peganum harmala L. seeds. Ultraperformance liquid chromatography–electrospray ionization–tandem mass spectrometry (UPLC/ESI-MS) analysis of HARF revealed 15 alkaloids. The reversed-phase high-performance liquid chromatography (RP-HPLC) analysis revealed three peaks: peganine, harmol, and harmine. The HARF was then encapsulated in p-SC6 nanocapsules employing a thin-film hydration approach. The designed nanocapsules had an average particle size of 264.8 ± 10.6 nm, and a surface charge of −30.3 ± 2.2 mV. They were able to encapsulate 89.3 ± 1.4, 74.4 ± 1.3, and 76.1 ± 1.7% of the three harmala alkaloids; harmine, harmol, and peganine; respectively. The in vitro drug release experiments showed the potential of the designed nanocapsules to release their cargo at a pH of 5.5 (typical of cancerous tissue). The IC50 values of HARF encapsulated in p-SC6 (H/p-SC6 nanocapsules) were 5 and 2.7 μg/mL against ovarian cancer cells (SKOV-3) and breast adenocarcinoma cells (MCF-7), respectively. The prepared nanocapsules were found to be biocompatible when tested on human skin fibroblasts. Additionally, the antioxidant activity of the designed nanocapsules was 5 times that of the free powder fraction; the IC50 of the H/p-SC6 nanocapsules was 30.1 ± 1.3 μg/mL, and that of the HARF was 169.3 ± 7.2 μg/mL. In conclusion, encapsulation of P. harmala alkaloid-rich fraction into self-assembled p-SC6 significantly increases its antioxidant and cytotoxic activities.